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Field distribution and activity of chlorinated solvents degrading bacteria by combining CARD-FISH and real time PCR

Paper ID Volume ID Publish Year Pages File Format Full-Text
33504 44981 2012 10 PDF Available
Title
Field distribution and activity of chlorinated solvents degrading bacteria by combining CARD-FISH and real time PCR
Abstract

Nowadays several advanced molecular techniques are applied for quantifying bacteria involved in contaminant degradation processes. However, despite the fact that significant efforts have been taken to make these tools more reliable and specific, their application for the analysis of field samples is hardly ever applied.In this study, a combination of three methods (CARD-FISH, qPCR and RT-qPCR) was successfully applied to evaluate the distribution and the activity of known chlorinated solvent dechlorinating bacteria in a contaminated site where no remedial actions have been undertaken.CAtalysed Reporter Deposition Fluorescence In Situ Hybridization (CARD-FISH) specifically provided the cell densities of known dechlorinating bacteria and was found to be more sensitive than quantitative PCR (qPCR) for the quantification of ‘Dehalococcoides’ cell numbers in the aquifer. Among the screened dechlorinators, ‘Dehalococcoides’ spp. were mainly found and nearly homogenously distributed in the aquifers at concentrations ranging from 8.1 × 105 ± 1.2 × 105 to 2.5 × 107 ± 5.6 × 106 cells per liter of groundwater (with a relative abundance out of the total Bacteria of 0.7–15%). Further, the dechlorination potentialities of ‘Dehalococcoides’ species living in the aquifer were evaluated by analyzing the abundance and the expression of 16S rRNA genes and reductive dehalogenase (RDase) encoding functional genes by qPCR and Reverse Transcription qPCR (RT-qPCR). ‘Dehalococcoides’ tceA gene, known to be associated to strains capable of reducing chlorinated solvents beyond cis-DCE, was found and expressed in the field. Overall, this study proved the existence of a well-established dechlorinating microbial community able to use contaminants as substrates for their metabolic activity and indicated the occurrence of reductive dechlorination at the site.

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Field distribution and activity of chlorinated solvents degrading bacteria by combining CARD-FISH and real time PCR
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Publisher
Database: Elsevier - ScienceDirect
Journal: New Biotechnology - Volume 30, Issue 1, 15 November 2012, Pages 23–32
Authors
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Subjects
Physical Sciences and Engineering Chemical Engineering Bioengineering
Get Full-Text Now
Don't Miss Today's Special Offer
Price was $35.95
You save - $31
Price after discount Only $4.95
100% Money Back Guarantee
Full-text PDF Download
Online Support
Any Questions? feel free to contact us